Objective To screen the conservative,stable and specific DNA signature sequence in the plasmid of Yersinia pestis.Methods Specific validation trials and stability of the qualification test were carried out to 40 strains of Yersinia pestis,47 strains of non-Yersinia pestis of home and wild types of rodent in Yunnan,by using 32 DNA sequences derived from Yersinia pestis in the plasmid and conventional PCR technology,and Yersinia pestis vaccine strain EV76 as a positive control.Results Four pairs of relatively conservative,stable and specific DNA marker genes were screened:YPMT1.05c,YPMT1.03c,YPMT1.42 and YPMT1.04c.Conclusions The 4 pairs of Yersinia pestis DNA signature sequences can be used for rapid diagnosis of plague.

Objective To explore the feasibility of culturing dermal papillae cells (DPC) of hu- man hair in a serum-flee medium,and to observe the growth characteristics of these cells.Methods Cell culture flasks (plates) were pretreated with fibronectin,and DPC (2nd passage) were incubated with Williams E serum-flee medium supplemented with insulin-transferrin-selenite (ITS).Cells were observed by an inverted phase-contrast microscope.Proliferation of DPC was evaluated with 3-(4,5-dimethylthia- zol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and by their growth curve.Results In a serum-free medium,2nd passage DPC adhered to the flask surface within two to four hours of incubation; two to three days later,confluence,of the cells was observed,without noticeable proliferation.Four days later,cell connection was interrupted,isolated cells or cell clusters were seen,and detachment of some cells from the flask surface was observed.One to two weeks later,most cells had died.After incubation with 4% bovine serum for ten hours,cell proliferation was observed surrounding the remaining viable cell colonies. DPC growth curve showed stagnant phase and slow growth phase;however,log growth phase was not ob- served.Conclusion DPC could be successfully cultured in serum-free medium.However,the culture con- dition needs to be further optimized.

Objective To compare the difference of biochemical characteristics and virulent Pst Ⅰ of Yersinia pestis strains in traditional focuses of plague in Yunna Province and in the new focuses of plague in Yulong County. Methods The identification data of biochemical characteristics(Rhamnose, Glycerol, Maltose, L-Arabina and Melibiose fermentation) and virulence factor(Pst Ⅰ) from different focuses of plague in Yunna Province were Retrospectively collected by tube test followed by the analysis using statistics software SAS 8.0 by Fisher exact probability of disordered two-way R × C table χ2 test. Results Among 48 strains of Yersinia pestis from hantaan type plague focus, 1 strain fermented L-maltose, 48 strains fermented Glycerol. Among 165 strains of Yersinia pestis from the Soul type plague focus, 1 strain did not ferment L-maltose, only one of them fermented Glycerol. 1 strain from the Soul type plague focus was confirmed to have mutation, for the test of nitrate reduction reaction was negative. All 5 strains of Yersinia pestis from the new focuses of plague in Yulong County fermented L-maltose and Glycerol. The statistical result showed that the differences in L-maltose and Glycerol fermentation of Yersinia pestis from different natural focuses of plague in Yunnan Province were statistically siguificant (P < 0.01). The differences of other biochemical characteristics and Pst Ⅰ were not statistically significant (P > 0.01). Conclusions Biochemical characteristics of Yersinia pestis from the hantaan type plague focus and the Soul type plague focus in Yunnan province are overlapping. Biochemical characteristics of Yersinia pestis from the new focuses of plague in Yulong County are different from those tradition focuses of plague in Yunna Province but share similarities to those from Unquiculatus focuses in North Tibet.

Antibodies, Monoclonal, Murine-Derived ; metabolism ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Hemangioendothelioma ; metabolism ; pathology ; surgery ; Hemangioendothelioma, Epithelioid ; metabolism ; pathology ; surgery ; Humans ; Lymphatic Metastasis ; Middle Aged ; Platelet Endothelial Cell Adhesion Molecule-1 ; metabolism ; Sarcoma ; metabolism ; pathology ; surgery ; Thigh ; Vimentin ; metabolism ; von Willebrand Factor ; metabolism

Adult ; Antiphospholipid Syndrome ; diagnosis ; Cesarean Section ; Diagnostic Errors ; Female ; Humans ; Infarction ; diagnosis ; Liver Diseases ; diagnosis ; Pregnancy

Objective To assese the healing of stoma after magnetic anastomosis for the reconstruction of biliary-enteric continuity under severe inflammation.
Methods Acute bile duct injury was constructed as a bile peritonitis model in mongrel dogs (n=32). Magnetic anastomosis (group A, n=16) and traditional suture anastomosis (group B, n=16) were performed to reconstruct the biliary-enteric continuity in one stage. Half of the dogs in each group were euthanized on the 30th postoperative day, and the other half on the 90th postoperative day to harvest the stoma region. The healing conditions of the stoma after the 2 anastomotic approaches were observed with naked eyes, under light microscope and scanning electron microscope.
Results The stoma leakage rate (50%versus 0%on the 30th postoperative day, 37.5%versus 12.5%on the 90th postoperative day, both P<0.05) and stenosis degree (13.9%±0.3%versus 7.1%±0.3%on the 30th postoperative day, 17.2%±0.4%versus 9.4%±0.4%on the 90th postoperative day, both P<0.01) were significantly higher in group B than in group A. Compared with traditional manual anastomoses, the histological analysis under light and electron microscope showed a more continuous stoma with more regular epithelium proliferation and collagen arrangement, less inflammation in group A.
Conclusions Magnetic anastomosis stent ensures better healing of the stoma even under the circumstance of severe inflammation.

OBJECTIVETo explore the use of the urinary neutrophil gelatinase associated lipocalin (uNGAL) in the early diagnosis of paraquat poisoning patients with acute kidney injury (AKI).

METHODSEighty five patients were from the emergency department in our hospital. Five ml blood and urine were collected from each patient at 15 min, 2, 4, 6, 8, 10, 12, 18, 24, 36, 48 and 72 h, 5 and 7d after admission. The uNGAL levels of urine were detected with ELISA test and the SCr levels were measured with creatine oxidase assay.

RESULTSSixty two cases of paraquat intoxication suffered from AKI, the incidence was 72.94% (62/85). The SCr levels of 62 cases with AKI at 18, 24, 36, 48, 72 h and 5, 7 d after admission increased significantly, as compared with the baseline value and control group (P < 0.01). At 24, 36, 48, 72 h and 5, 7 d after admission, there was significant difference of the SCr levels between AKI group and non-AKI group (P < 0.01). At 2 h after admission, the uNGAL level of urine in paraquat intoxication AKI group was (96.21 +/- 45.32) microg/L which was significantly higher than the baseline value. At 10, 12, 18, 24, 36, 48, 72 h and 5, 7 d after admission, the uNGAL levels of urine in AKI group and non-AKI group obviously enhanced, as compared with the baseline value and control group (P < 0.01 or P < 0.05). At all time points, there was significant difference of the uNGAL level between AKI group and non-AKI group (P < 0.01).

CONCLUSIONThe uNGAL level of urine in paraquat intoxication patients at 2 h after admission significantly enhanced, which is earlier than enhanced SCr. So the uNGAL level of urine may serve as early diagnostic biomarker for AKI induced by paraquat intoxication.

Acute Kidney Injury ; chemically induced ; diagnosis ; Acute-Phase Proteins ; urine ; Adolescent ; Adult ; Case-Control Studies ; Early Diagnosis ; Female ; Humans ; Lipocalin-2 ; Lipocalins ; urine ; Male ; Middle Aged ; Paraquat ; poisoning ; Proto-Oncogene Proteins ; urine ; Young Adult

OBJECTIVETo investigate the relations between anti-myelin basic protein antibody (anti-MBP) variation and myelinoclasis in the brain stem following brain trauma.

METHODSIn rat models of brain trauma, MBP content and anti-MBP titer in the blood were measured using enzyme-linked immunosorbent assay (ELISA) at different time points after brain trauma, and the degree of myelinoclasis in the brain stem slices was assessed with osmic acid staining.

RESULTSEarly after brain trauma, MBP content in the blood increased followed by significant reduction 10 days later. Four days after the trauma, anti-MBP titer was markedly increased, accompanied by obvious exacerbation of myelinoclasis in the brain stem, both reaching the highest levels on day 10, at the point of which anti-MBP titer increased by 4 folds and the number of myelinoclasis by 10 folds compared with the control group. Anti-MBP titer and brain stem myelinolysis both lowered 30 days later. Correlation analysis showed an intimate positive correlation between anti-MBP titer and the degree of myelinoclasis.

CONCLUSIONAfter brain trauma, MBP is released as a specific antigen into the blood to stimulate the immune system for anti-MBP production, and the antibody is intimately related to the brain stem myelinoclasis.

Animals ; Antibodies ; metabolism ; Brain Injuries ; complications ; Brain Stem ; immunology ; pathology ; Demyelinating Autoimmune Diseases, CNS ; etiology ; immunology ; Female ; Male ; Myelin Basic Protein ; Nerve Tissue Proteins ; blood ; immunology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Transcription Factors ; blood ; immunology

OBJECTIVETo investigate the change of inflammatory factor in lung tissue of acute paraquat (PQ) poisoned rats.

METHODShundred SD rats were randomly divided into two groups: the normal control group (NC group, n = 10) and the PQ group (n = 80). The 1 ml saline was administered once in normal control group. The PQ group was administered with 25 mg/kg 1% PQ by intraperitoneal injection to establish the model of PQ induced renal injury. At six hours, at the first, the third and the seventh day the PQ group were sacrificed, while at the first day the normal control group was sacrificed. The level of normal tumor necrosis factor-α (TNF-α), interleukin-2 (IL-2), IL-6 in serum of rats were detected. Meanwhile, pathological changes of the renal were examined under optical microscope.

RESULTSHistopathological findings of an earlier, a large number of patients edema clearly inflammatory cell infiltration. Compared with the control group, PQ exposure of serum TNF-α, IL-2, IL-6, the level at each time point were elevated. PQ treated group 6 h and 1, 3, 7 d when the IL-2 levels were (2.16 ± 0.65), (2.95 ± 1.02), (3.05 ± 1.12), (2.21 ± 0.62) µg/L, IL-6 were (62.5 ± 8.6), (85.6 ± 13.5), (90.3 ± 15.6), (65.3 ± 9.1) ng/ml, TNF-α were (1.95 ± 0.53), (2.86 ± 0.92), (3.15 ± 1.02), (2.06 ± 0.71) µg/L, compared with the control group, are significantly higher, the differences were statistically significant (P < 0.01).

CONCLUSIONacute PQ poisoning serum TNF-α, IL-2, IL-6 levels were significantly increased both early and late inflammatory factors involved in PQ poisoning the pathogenesis of renal injury.

Animals ; Interleukin-2 ; blood ; Interleukin-6 ; blood ; Kidney ; pathology ; Male ; Paraquat ; poisoning ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo investigate the expression dynamics and significance of matrix metalloproteinase-2 (MMP-2) membrane type-matrix metalloproteinase-2 (MT-MMP-2) in hepatic fibrosis and its reversal counterpart.

METHODSAn experimental CCl4 induced hepatic fibrosis rat model was established by intraperitoneal administration of carbon tetrachloride for 2, 4, 6, 8, 10 weeks, and normal rats were used as a control group. The immunohistochemical methods and in situ hybridization were used to detect MMP-2,MT-MMP-2 mRNA and related antigens in the liver.

RESULTSMMP-2,MT-MMP-2 mRNA and related antigens were expressed in mesenchymal cells and parts of hepatocytes besides active pathological changes, especially in the fibrous septum and portal area. Expression of MMP-2,MT-MMP-2 mRNA and related antigens were increased in hepatic fibrosis and decreased gradually in its reversal counterpart.

CONCLUSIONThis study suggested that mesenchymal cells are the main cellular origins of MMPs. The levels of MMP-2 and MT-MMP-2 antigens and gene expression were closely related to hepatic fibrosis. MMP-2 and MT-MMP-2 may play important roles in hepatic fibrosis and its reversal counterpart.

Animals ; Carbon Tetrachloride Poisoning ; Gene Expression Regulation, Enzymologic ; Hepatocytes ; enzymology ; Liver ; enzymology ; pathology ; Liver Cirrhosis, Experimental ; enzymology ; etiology ; pathology ; Male ; Matrix Metalloproteinase 2 ; biosynthesis ; genetics ; Matrix Metalloproteinases ; biosynthesis ; genetics ; Matrix Metalloproteinases, Membrane-Associated ; Mesenchymal Stromal Cells ; enzymology ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Wistar